Info - Leica Biosystems

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PA0171 Rev A
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Minimize microbial contamination of reagents or an increase in non-specific staining may occur.
Retrieval, incubation times or temperatures other than those specified may give erroneous results. Any such change must be
validated by the user.
Instructions for Use
CD21 (2G9) primary antibody was developed for use on the automated Bond system in combination with Bond Polymer Refine
Detection. The recommended staining protocol for CD21 (2G9) primary antibody is IHC Protocol F. Heat induced epitope retrieval is
recommended using Bond Epitope Retrieval Solution 2 for 20 minutes.
Results Expected
Normal Tissues
Clone 2G9 stained the CD21 protein in follicular dendritic cells (FDCs) of lymphoid tissue and mature B cells. It strongly labelled FDCs
in 29/29 cases of tonsil, with a dense meshed FDC network in the light zone and a more loosely arranged, weaker staining pattern of
staining in the dark zone. The staining of mantle zone B cells was faint and much less consistent. The staining of the FDC network was
also seen in 5/5 cases of spleen. FDC staining was also evident in 2/4 cases of thyroid, 2/2 cases of cervix, 3/4 cases of stomach and
3/4 cases of colon. The occasional mononuclear cell was labelled in other normal tissues tested. There was some weak staining of
Leydig cells in testis no additional staining was observed in cases of uterus, placenta, prostate, ovary, pancreas, lung, liver, skin, breast,
heart, small intestine, brain, pituitary and adrenal (n = 200).
Tumor Tissues
With clone 2G9, in follicular lymphoma (n = 3) the atretic follicles exhibited strong, well-defined FDC networks. In the larger abnormal
follicles a more diffuse and less intense staining of FDCs was observed. No expression of CD21 was observed in the neoplastic cells
of T-cell lymphoma (n=6) other than a few residual follicles showing strong FDC staining. No staining of neoplastic cells was observed
in non-Hodgkin’s B-cell lymphoma (n = 14), lymphoblastic lymphoma (n = 2), Hodgkin’s disease (n = 4) or a range of additional tumors
(n=26).
CD21 (2G9) is recommended for the immunodetection of normal and abnormal cell expression of CD21 in follicular dendritic cells.
Product Specific Limitations
CD21 (2G9) has been optimized at Leica Biosystems for use with Bond Polymer Refine Detection and Bond ancillary reagents.
Users who deviate from recommended test procedures must accept responsibility for interpretation of patient results under these
circumstances. The protocol times may vary, due to variation in tissue fixation and the effectiveness of antigen enhancement, and must
be determined empirically. Negative reagent controls should be used when optimizing retrieval conditions and protocol times.
Troubleshooting
Refer to reference 3 for remedial action.
Contact your local distributor or the regional office of Leica Biosystems to report unusual staining.
Further Information
Further information on immunostaining with Bond reagents, under the headings Principle of the Procedure, Materials Required,
Specimen Preparation, Quality Control, Assay Verification, Interpretation of Staining, Key to Symbols on Labels, and General Limitations
can be found in “Using Bond Reagents” in your Bond user documentation.
Bibliography
1. Clinical Laboratory Improvement Amendments of 1988, Final Rule 57 FR 7163 February 28, 1992.
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Villanova PA. National Committee for Clinical Laboratory Standards (NCCLS). Protection of laboratory workers from infectious
diseases transmitted by blood and tissue; proposed guideline. 1991; 7(9). Order code M29-P.
Bancroft JD and Stevens A. Theory and Practice of Histological Techniques. 4th Edition. Churchill Livingstone, New York. 1996.
Kojima M, Nakamura S, Shimizu K, et al. Classical Hodgkin lymphoma occurring in clusters of nodal marginal zone
B-lymphocytes in association with progressive transformation of germinal center. Pathology Research and Practice. 2003;
199:547–550.
Kojima M, Nakamura S, Motoori T, et al. Primary marginal zone B-cell lymphoma of the lymph node resembling plasmacytoma
arising from a plasma cell variant of Castleman’s disease. A clinicopathological and immunohistochemical study of seven patients.
Acta Pathologica, Microbiologica et Immunologica Scandinavica (APMIS). 2002; 110:875–880.
Dupin N, Fisher C, Kellam P et al. Distribution of human herpesvirus-8 latently infected cells in Kaposi’s sarcoma, multicentric
Castleman’s disease, and primary effusion lymphoma. Proceedings of the National Academy of Sciences USA. 1999; 96(8):
4546–4551.
ProClin 950 is a trademark of Supelco, a part of Sigma-Aldrich Corporation.
Date of Issue
17 April 2008